Fig. 3
Circ-YAP encodes YAP-220aa mediated by m6A. A, B. The sketch and full-length sequence of circ-YAP. C. The sketch showing the construction of the indicated vectors. D. Western blot analysis of Flag and YAP-220aa protein levels in 293T cells transfected with the above vectors. E. Coomassie blue staining of protein samples from control and circ-YAP-overexpressing cells, followed by mass spectrometry of the indicated gels. F. Western blot analysis of YAP-220aa protein levels in circ-YAP-overexpressing cells. G. IF staining of Flag and YAP-220aa in 293T cells. Scale bar, 25 μm. H. Western blot analysis of YAP-220aa protein levels in CRC cell lines. I. Western blot testing the effect of METTL3 knockdown on YAP-220aa expression. J. meRIP assay testing the m6A levels on circ-YAP in CRC cells. K, L. Cell migration and invasion in HT29 and SW480 cells transfected with the indicated vectors. M. The experimental liver metastasis model testing the effect of circ-YAP or circ-YAP-m6A-mutation on CRC liver metastasis (n = 5 per group). ***P < 0.001. Data (J, K, L) are the mean ± SD of three independent experiments carried out in triplicate. The uncropped western blot data are provided as a Original Blot Image file